Gene delivery vectors with cell type specificity for primary human chondrocytes

Abstract

The present invention relates to a gene delivery vehicle comprising a recombinant adenovirus having a tropism for a primary human chondrocyte. By efficiently transducing a nucleic acid of interest into a primary chondrocytes, the gene delivery vehicle is able to at least in part improve the counteraction of cartilage disease. In one embodiment the recombinant adenovirus comprises a deletion in the gene encoding for fiber protein, which is replaced by a nucleic acid sequence encoding at least part of a fiber protein of a B-type adenovirus.

What is claimed is:  
     
       1. An in vitro method of delivering a nucleic acid of interest to a primary human chondrocyte, comprising: 
       providing a recombinant adenovirus having a tropism for primary human chondrocytes, said recombinant adenovirus comprising:  
       a nucleic acid of interest operatively linked to a promoter, wherein said nucleic acid of interest encodes at least one amino acid sequence that inhibits cartilage disease progression, at least one amino acid sequence that counteracts the loss of cartilage, or a combination thereof;  
       a deletion in a gene encoding a fiber protein; and  
       a nucleic acid replacing the deletion in the gene of the fiber protein, said nucleic acid encoding at least a part of a fiber protein of a B-type adenovirus, and wherein said at least a part of the fiber protein of the B-type adenovirus has a tropism for primary human chondrocytes; and  
       infecting a primary human chondrocyte in vitro with said recombinant adenovirus, such that said nucleic acid of interest is delivered to said primary human chondrocyte.  
     
     
       2. The method of  claim 1 , wherein said B-type adenovirus is adenovirus type 35. 
     
     
       3. The method of  claim 1 , wherein said recombinant adenovirus comprises an adenovirus 5 nucleic acid sequence. 
     
     
       4. The method of  claim 2 , wherein said recombinant adenovirus comprises an adenovirus 5 genome. 
     
     
       5. The method of  claim 1 , wherein said recombinant adenovirus comprises at least one deletion in the E3 region where the nucleic acid of interest is inserted or can be inserted. 
     
     
       6. In vitro chondrocytes provided with an additional nucleic acid encoding: 
       at least one amino acid sequence that inhibits cartilage disease progression;  
       at least one amino acid sequence that counteracts the loss of cartilage; or  
       a combination thereof;  
       said additional nucleic acid provided by a gene delivery vehicle comprising a recombinant adenovirus having a tropism for chondrocytes;  
       said recombinant adenovirus comprising:  
       a deletion in a gene encoding a fiber protein; and  
       a nucleic acid replacing the deletion in the gene encoding the fiber protein, said nucleic acid encoding at least a part of a fiber protein of a B-type adenovirus  
       wherein said at least a part of the fiber protein of the B-type adenovirus has a tropism for primary human chondroctyes.  
     
     
       7. The in vitro chondrocytes of  claim 6 , wherein said additional nucleic acid encodes at least one member of the family of bone morphogenesis proteins. 
     
     
       8. An in vitro method of transducing a primary human chondrocyte, the method comprising: 
       preparing a recombinant adenovirus having a tropism for primary human chondrocytes, said recombinant adenovirus including:  
       a nucleic acid encoding a protein useful in inhibiting cartilage disease progression operatively linked to a promoter;  
       a deletion in a gene encoding a fiber protein; and  
       a nucleic acid replacing the deletion in the gene encoding the fiber protein, said nucleic acid encoding at least a part of a fiber protein of a B-type adenovirus, and wherein said at least a part of the fiber protein of the B-type adenovirus has a tropism for primary human chondrocytes; and  
       infecting a primary human chondrocyte in vitro with said recombinant adenovirus, such that said nucleic acid of interest encoding the protein useful in inhibiting cartilage disease progression is expressed in said primary human chondrocyte.  
     
     
       9. An in vitro method of transducing a primary human chondrocyte, the method comprising: 
       preparing a recombinant adenovirus having a tropism for primary human chondrocytes, said recombinant adenovirus including:  
       a nucleic acid encoding a protein useful in repairing cartilage operatively linked to a promoter;  
       a deletion in a gene encoding a fiber protein; and  
       a nucleic acid replacing the deletion in the gene encoding the fiber protein, said nucleic acid encoding at least a part of a fiber protein of a B-type adenovirus, and wherein said at least a part of the fiber protein of the B-type adenovirus has a tropism for primary human chondrocytes; and  
       infecting a primary human chondrocyte in vitro with said recombinant adenovirus, such that said nucleic acid encoding the protein useful in repairing cartilage is expressed in said primary human chondrocyte.  
     
     
       10. An in vitro method of delivering a nucleic acid of interest to a primary human chondrocyte, comprising: 
       providing a recombinant adenovirus having a tropism for primary human chondrocytes, said recombinant adenovirus comprising:  
       a nucleic acid of interest operatively linked to a promoter, wherein said nucleic acid of interest encodes at least one member of the family of bone morphogenesis proteins;  
       a deletion in a gene encoding a fiber protein; and  
       a nucleic acid replacing the deletion in the gene of the fiber protein, said nucleic acid encoding at least a part of the fiber protein of a B-type adenovirus, wherein said at least a part of the fiber protein of the B-type adenovirus has a tropism for primary human chondrocytes; and  
       infecting a primary human chondrocyte in vitro with said recombinant adenovirus, such that said nucleic acid of interest is delivered to said primary human chondrocyte.